Biomarker Approach to Screening for the early detection of HPV-related Oropharyngeal Cancer (BASH OPC)

摘要 口咽癌(OPC)的发病率正在显著增加，大多数病例(~80%)是由人类引起的 乳头瘤病毒(HPV)感染16型。大多数OPC病例诊断较晚，需要加强治疗 化学辐射导致严重的发病率、终生残疾和死亡。早期的OPC检测目前 最可行的二级预防方法，有可能极大地改善OPC患者的预后。 我们完成了一系列研究，重点是开发一种适用于早期OPC的口腔漱口生物标志物小组 可在单个口腔含漱液样本中测量的检测，并可区分早期OPC(T1- 2例N0-1[同侧单个小肿瘤结节≤为3 cm]病例组和对照组，设两组病例对照。 在一项后续研究中，生物标记物小组的特异性和敏感性得到了提高(AuC=0.935)，基因组- 广泛的甲基化阵列发现方法。这导致小组扩大到包括口服HPV16状态和 13个差异甲基化宿主基因CpG位点。我们现在准备推进这个口腔漱口生物标志物小组 到生物标记物开发的第二阶段。我们建议利用早期OPC的现有生物资源库 两个癌症中心的病例和研究基础设施，以有效地推动临床分析开发和 验证。我们的目标是验证一种非侵入性诊断生物标记物小组，以早期发现OPC。中环 假设是，我们可以使用与以下相关的生物标志物来区分早期OPC病例和未患癌症的患者 HPV和宿主表观遗传学改变，从而识别可以有效和安全地使用 存活率最高的单一模式。由于我们敏锐地意识到这是一个不断发展的领域，我们还将 随着文献的发展，对这些和其他生物标志物进行初步评估。其主要目的是 联合检测HPV16DNA和宿主基因甲基化口服生物标记物的敏感性和特异性 在100例早期和100例晚期疾病治疗前的OPC中区分早期OPC和对照组的小组 来自莫菲特癌症中心和UPMC的病例和200名按性别、年龄、种族和烟草匹配的对照 希尔曼癌症中心。次要目标包括评估实验室间和实验室内生物标记物的一致性 和可靠性，在病例和对照中分别与生物标记物小组相关的因素，以及 其他差异甲基化宿主基因(例如SYNGR3)和/或口服肿瘤组织修饰的HPV的性能 DNA以早期发现OPC。这是一种使用非侵入性标本进行OPC早期检测的创新方法 容易获得的(例如，在常规的牙科实践中)。该方法建立在现有的临床/流行病学基础上。 基础设施、注释良好的生物储存库和来自OPC案例和控制的数据，以及强大的初步数据。 我们的研究团队拥有成功实施拟议研究的经验和专业知识。结果将 告知在非侵入性试验中测量的生物标志物小组的大型确定性前瞻性筛查试验的设计 在高危人群中采集样本，将OPC的诊断从晚期转移到早期。

ABSTRACT Oropharyngeal cancer (OPC) incidence is significantly increasing with most cases (~80%) caused by human papillomavirus (HPV) infection type 16. The majority of OPC cases are diagnosed late and require intensive chemo-radiation causing significant morbidity, life-long disabilities, and mortality. Early OPC detection is currently the most viable secondary prevention approach with the potential to vastly improve outcomes for OPC patients. We completed a series of studies focused on developing an oral gargle biomarker panel suitable for early OPC detection that can be measured in a single oral gargle specimen, and can discriminate between early OPC (T1- 2 N0-1 [small tumors with a single ipsilateral node ≤3 cm]) cases and controls in two different case-control sets. In a follow-up study the biomarker panel specificity and sensitivity was improved (AUC = 0.935) with a genome- wide methylation array discovery approach. This resulted in panel expansion to include oral HPV 16 status and 13 differentially methylated host gene CpG sites. We are now poised to advance this oral gargle biomarker panel to the second phase of biomarker development. We propose to leverage existing biorepositories of early OPC cases and research infrastructure at two cancer centers to efficiently advance clinical assay development and validation. Our goal is to validate a non-invasive diagnostic biomarker panel to detect OPC early. The central hypothesis is that we can distinguish early OPC cases from cancer-free individuals using biomarkers related to HPV and host epigenetic alterations, thereby identifying tumors that can be effectively and safely treated with a single modality where survival is highest. As we are keenly aware that this is an evolving field, we will also conduct preliminary evaluation of these and other biomarkers as the literature evolves. The Primary Aim is to estimate the sensitivity and specificity of a combined HPV 16 DNA and host gene methylation oral biomarker panel to distinguish early OPC cases from controls among 100 early and 100 late disease pre-treatment OPC cases, and 200 controls matched by sex, age, race, and tobacco from the Moffitt Cancer Center and the UPMC Hillman Cancer Center. Secondary Aims include assessing inter- and within-laboratory biomarker concordance and reliability, factors associated with the biomarker panel separately among cases and controls, and the performance of other differentially methylated host genes (e.g., SYNGR3) and/or oral tumor-tissue modified HPV DNA to detect OPC early. This is an innovative approach to OPC early detection, using a non-invasive specimen readily obtained (e.g., in routine dental practice). The approach builds on existing clinical/epidemiologic infrastructure, well annotated biorepository and data from OPC cases and controls, and strong preliminary data. Our research team has the experience and expertise to successfully implement the proposed study. Results will inform the design of a large definitive prospective screening trial of a biomarker panel measured in a non-invasive specimen among high-risk individuals to shift OPC diagnosis from late to early disease.